摘要
目的 观察臭氧氧化预处理对大鼠肾脏缺血再灌注损伤导致的细胞凋亡的影响.方法 分为3组进行实验:(1)假手术组:切除大鼠右肾,缝合腹壁;(2)缺血再灌注组:切除大鼠右肾后,夹闭左肾动、静脉45 min,然后开放;(3)臭氧氧化预处理组:手术步骤与缺血再灌注组相同,在术前15d开始经直肠吹入氧气和臭氧的混合气体5~5.5 ml(臭氧浓度为50 mg/L,1 mg·kg-1 ·d-1),应用至术前1d.全自动生化分析仪检测3组大鼠血清尿素氮和肌酐,比色法测定血清脂质过氧化产物丙二醛(MDA)和超氧化物歧化酶(SOD).免疫组织化学法检测大鼠肾细胞胞浆细胞色素C(CytC)的表达,蛋白质印迹法检测CytC的含量.逆转录聚合酶链反应法检测肿瘤坏死因子α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素-6 (IL-6) mRNA的表达.结果 缺血再灌注和臭氧氧化预处理组血清尿素氮、肌酐和MDA均高于假手术组,而缺血再灌注组的尿素氮、肌酐和MDA又高于臭氧氧化预处理组.缺血再灌注组血清SOD低于假手术组和臭氧氧化预处理组(P<0.05).臭氧氧化预处理组的肾组织病理学改变较缺血再灌注组轻.假手术组、缺血再灌注组和臭氧氧化预处理组的CytC表达灰度值分别为101.50±18.02、181.00±20.85和156.71±16.82,两两比较,差异均有统计学意义(P<0.05).臭氧氧化预处理组肾细胞胞浆和线粒体中CytC含量较缺血再灌注组有所下降(P<0.05).结论 臭氧氧化预处理可以减轻肾脏脂质过氧化反应,减少炎症因子的合成,抑制CytC的释放,减轻肾缺血再灌注损伤.
Objective To investigate the protective effect of ozone oxidative preconditioning (OzoneOP) on apoptosis induced by acute renal ischemia/reperfusion (I/R) injury in rats.Methods The right kidneys of rats in control group were excised.A rat I/R model was established in I/R group.In OzoneOP group,the renal OzoneOP was induced by rectal insufflation of 5.0~5.5 ml oxygen plus ozone (ozone 50 mg/L,1 mg· kg- 1 · d- 1,once every day).The parameters (blood urea nitrogen and creatine ) of renal function were determined by auto-biochemical analyzer. The concentrations of malondialdehyde (MDA) and superoxide dismutase (SOD) were measured by using chromometry.Cytochrome c (CytC) was examined by using immunohistochemistry and Western blotting.The mRNA expression levels of TNF-α,IL-1β and IL-6 in the renal tissue were detected by using RT-PCR.Results Compared with the control group,the blood urea nitrogen,creatine,MDA and the release of CytC were increased significantly in I/R group.After OzoneOP,the levels of blood urea nitrogen,creatine,MDA,CytC and the mRNA expression levels of TNF-α,IL-1-β and IL-6were significantly decreased and SOD levels were significantly increased.Conclusion OzoneOP could relieve renal I/R injury of rats by improving antioxidation capability,cutting down cytokines contents and inhibiting the release of CytC from mitochondria.
出处
《中华器官移植杂志》
CAS
CSCD
北大核心
2012年第2期113-117,共5页
Chinese Journal of Organ Transplantation
基金
基金项目:国家自然科学基金(30901552,30901494)
关键词
大鼠
肾
臭氧
氧化预处理
再灌注损伤
细胞凋亡
Rats
Kidney
Ozone
Oxidative preconditioning
Reperfusion injury
Apoptosis